Australia's final opening round match at the Yokohama Baseball Stadium was basically a playoff to face Canada in Tuesday's bronze medal softball game.
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Jade Wall hit a consolation home run as Australia were punished for leaving five runners on base in the first three innings, going down 4-1 to finish fifth in the six-team competition.
Mexico took control after Suzannah Brookshire's second inning strike ricocheted off the wall to her advantage.
Brookshire's triple allowed Brittany Cervantes to score and she soon completed a run herself to double Mexico's lead.
Australia pitcher Kaia Parnaby was taken out of the game after Victoria Vidales' double in the fourth innings saw Tatyana Forbes scamper home.
New pitcher Gabrielle Plain could not stop Vidales scoring and Australia, who had won only one of their previous four games, were staring defeat in the face again.
Wall's clean strike in the sixth inning gave Australia brief hope, but impressive pitcher Dallas Escobedo closed out victory for Mexico.
Australia had won a medal at each of softball's four previous Olympics appearances between 1996 and 2008.
The United States and Japan will meet in Tuesday's gold medal match despite the hosts being beaten 2-1 by the Americans in their final opening round game on Monday.
Australian Associated Press
